BIND Chemistry for Imaging Intracellular Targets

用于细胞内靶标成像的 BIND 化学

基本信息

批准号：
8566732
负责人：
RALPH WEISSLEDER, MD, PHD
金额：
$ 11.47万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8566732
关键词：
A Mouse Affect Affinity Alder plant Amino Acids Appearance BODIPY Behavior Binding Binding Proteins Biochemical Biocompatible Biological Biological Models Cancer cell line Cell Culture Techniques Cell Nucleus Cells Centrosome Chemicals Chemistry Collaborations Combined Modality Therapy Coupled Data Detection Development Dose Drug Kinetics Drug resistance Enzymes Fluorescence Generic Drugs Genetic Engineering Goals Gold Housing Human Image Imagery In Vitro Interphase KRAS2 gene Kidney Kinetics Label Lead Libraries Life Ligands Liver Malignant Neoplasms Mass Spectrum Analysis Measures Methods Microscopic Mitosis Mitotic spindle Modeling Mus Organ PLK1 gene Pancreas Pancreatic Ductal Adenocarcinoma Pathway Analysis Patients Pharmaceutical Preparations Phosphotransferases Physicians Poly(ADP-ribose) Polymerases Positron-Emission Tomography Process Protein Binding Proteins Proteome Proteomics Reaction Reagent Recombinant Fusion Proteins Reporter Research Signal Transduction Small Interfering RNA Source Stable Isotope Labeling Technology Testing Therapeutic Toxic effect Treatment Efficacy Tumor Cell Line Work base cancer cell catalyst cellular imaging chemical synthesis comparative cycloaddition cytotoxic gemcitabine human PLK1 protein imaging modality imaging probe in vivo intravital imaging intravital microscopy molecular imaging mouse model novel protein complex research study response scaffold small molecule success therapeutic target tumor validation studies whole body imaging

项目摘要

There is a need to develop more sensitive, specific and robust strategies capable of imaging and quantitating therapeutic targets In cancers in vivo. To date, many biological targets and processes remain largely unobserved due to a) lack of affinity ligands, b) mismatches between target abundance and the amount of imaging agent required for target visualization, c) delivery barriers, d) unfavorable pharmacokinetics, e) high background signals, f) the unavailability of appropriate affinity ligands, and/or g) the sheer numbers of targets. The goal of this project is to use a recently developed bioorthogonal platform technology (known as BIND) for the rapid development of intracellular imaging agents such as polo-lil<e kinase (PLK1) and poly-ADP-ribose-polymerase 1 (PARP1), which are key intracellular cancer targets. BIND involves a {4+2} inverse Diels-Alder cycloaddition between fraA7s-cyclooctene (TCO) and a tetrazine (Tz), either attached to an imaging reporter or to a small molecule affinity ligand. BIND is biocompatible, has one of the most favorable reaction kinetics among bioorthogonal reactions, does not require catalysts, works well with small-molecule therapeutic drugs and can be combined with efficient amplifications strategies. This reaction can also be easily incorporated into development strategies for the vast number of existing small molecule affinity ligands for kinases and other intracellular targets. Cell permeable, small-molecule binders for PLK1 and PARP will be modified using TCO/Tz and imaged using complementary TCO/Tzimaging reporters. We will first validate the approach using cell based microscopic imaging, before performing in vivo experiments in mouse models of pancreatic ductal adenocarcinoma (PDAC). The specific aims are: 1) to develop a library of PLK-1 and PARP1-targeted BIND agents and characterize them biochemically, using live cell imaging and intravital microscopy; 2) to perform target-identification and network analysis using in vivo proteomics (SILAC); and 3) to explore the translational potential using ^^F-labeled agents in gemcitabine and PLKHnhibitor/PARPinhibitor treated mice.

需要开发更灵敏、更特异和更稳健的策略，能够成像和定量体内癌症的治疗靶点。迄今为止，许多生物靶标和过程由于 a) 缺乏亲和配体，b) 靶标丰度之间的不匹配，很大程度上仍未被观察到以及目标可视化所需的显像剂的量，c) 递送障碍，d) 不利药代动力学，e) 高背景信号，f) 缺乏适当的亲和配体，和/或 g) 目标的绝对数量。该项目的目标是使用最近开发的生物正交平台技术（称为 BIND）用于细胞内显像剂的快速开发，例如 polo-lil<e 激酶 (PLK1) 和聚 ADP 核糖聚合酶 1 (PARP1)，是细胞内癌症的关键靶点。 BIND 涉及到 fraA7s-环辛烯 (TCO) 和四嗪 (Tz) 之间的 {4+2} 逆狄尔斯-阿尔德环加成反应，连接到成像报告分子或小分子亲和配体。 BIND 具有生物相容性，具有以下之一生物正交反应中最有利的反应动力学，不需要催化剂，有效与小分子治疗药物配合良好，并且可以与有效的扩增策略相结合。这种反应也可以很容易地纳入大量现有的发展战略中。激酶和其他细胞内靶标的小分子亲和配体。细胞渗透性、小分子 PLK1 和 PARP 的结合物将使用 TCO/Tz 进行修饰，并使用互补的 TCO/Tzimaging 进行成像记者。在进行体内实验之前，我们将首先使用基于细胞的显微成像来验证该方法胰腺导管腺癌（PDAC）小鼠模型的实验。具体目标是：1）开发一个 PLK-1 和 PARP1 靶向 BIND 试剂库，并使用它们进行生化表征活细胞成像和活体显微镜检查； 2）使用以下方法进行目标识别和网络分析体内蛋白质组学（SILAC）； 3) 探索使用 ^^F 标记试剂的转化潜力吉西他滨和 PLKHnhibitor/PARPinhibitor 治疗的小鼠。